Anti-inflammatory Efficacy of

Emu Oil
Refinement of an in vitro assay

by Dr Christine A Lunam

January 2008
RIRDC Publication No 08/010
RIRDC Project No UF-13A

 2008 Rural Industries Research and Development Corporation.

All rights reserved.

ISBN 1 74151 599 8

ISSN 1440-6845
Anti-inflammatory efficacy of emu oil: refinement of an in vitro assay
Publication No. 08/010
Project No. UF-13A
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and to help improve the development of sustainable regions. You must not rely on any information contained in
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While reasonable care has been taken in preparing this publication to ensure that information is true and correct,
the Commonwealth of Australia gives no assurance as to the accuracy of any information in this publication.
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should be addressed to the RIRDC Publications Manager on phone 02 6271 4165

Researcher Contact Details

Dr Christine Lunam
Department of Anatomy & Histology
Flinders University
PO Box 2100
ADELAIDE SA 5001
Phone: 08 8204 4704
Fax:
08 8277 0085
Email: chris.lunam@flinders.edu.au
In submitting this report, the researcher has agreed to RIRDC publishing this material in its edited form.
RIRDC Contact Details
Rural Industries Research and Development Corporation
Level 2, 15 National Circuit
BARTON ACT 2600
PO Box 4776
KINGSTON ACT 2604
Phone: 02 6271 4100
Fax:
02 6271 4199
Email:
rirdc@rirdc.gov.au.
Web:
http://www.rirdc.gov.au

Published in January 2008

Printed by Canprint

ii

Foreword
The aim of this short term study is to refine an in vitro test to provide a quantitative measure of the
potential anti-inflammatory efficacy of emu oil. Subsequent work will then investigate factors that
affect the anti-inflammatory activity of emu oil using the in vitro assay as a measure of its potential
anti-inflammatory efficacy.
To maximise the market potential for emu oil, the Emu Industry needs to overcome two problems.
The first is to identify those factors that result in the different levels of its anti-inflammatory activity,
thereby allowing the consistent production of oil with known anti-inflammatory efficacy. The second
problem is to develop a sensitive in vitro assay to provide a quantitative measure of the potential antiinflammatory activity of each batch of oil.
This report describes the effect of emu oil on the in vitro production of inflammatory mediators from
stimulated human lymphocytes. This report also provides a methodology to form a stable emulsion of
emu oil that is non-toxic to human lymphocytes.
This project was funded from RIRDC Core Funds which are provided by the Australian Government
for the program area of New Animal Products.
This report, an addition to RIRDCs diverse range of over 1700 research publications, forms part of
our New Animal Products R&D program, which aims to accelerate the development of viable new
animal industries.
Most of our publications are available for viewing, downloading or purchasing online through our
website:
downloads at www.rirdc.gov.au/fullreports/index.html
purchases at www.rirdc.gov.au/eshop

Peter OBrien
Managing Director
Rural Industries Research and Development Corporation

iii

Acknowledgments

Dr Peter McInnes for his support and encouragement to undertake this study.

Mr Geoff Lean, (Industry Partner) for his enthusiasm, the fruitful discussions of the work and
hands-on assistance in preparation of the emulsified oil samples at Flinders University.

Dr Peter Macardle, Director of the Department of Immunology, Allergy and Arthritis at

Flinders Medical Centre for his advice on development of the in vitro assay and interpretation
of the data, for overseeing the assays, and for supplying the culture media and many other
reagents used in the preparation of the cultures.

Ms Rachel Hall for preparation of the cultures and conducting the cytokine assays. Thank you
also Rachel for supplying the information on the methodology and data generated from the
assays.

Mr Chris Gregory, (Industry Partner) managing director of Emu Tracks Australasia Pty Ltd
for supplying the emu oils used in this study.

Dr John Plummer, Chief Medical Scientist in the Department of Anaesthesia & Pain
Management at Flinders Medical Centre for his advice on the preparation of the oil samples.

Dr Michael Story (Industry Partner) for discussions regarding the initial preparation of the
emulsified oil samples and for supplying the cyclodextrins used in the initial oil preparations.

New Animal Products Research & Development Program, RIRDC, for funding the work.

iv

Contents
Foreword ............................................................................................................................................... iii
Acknowledgments................................................................................................................................. iv
Contents.................................................................................................................................................. v
Executive Summary ............................................................................................................................. vi
1.
Introduction................................................................................................................................. 1
1.1 Background and overall aims ............................................................................................. 1
1.2 Specific aims....................................................................................................................... 1
1.3 Emu oil industry perspective ........................................................................................... 1
2.
Methods........................................................................................................................................ 3
2.1 Emulsification of the emu oil.............................................................................................. 3
2.2 Lymphocyte source............................................................................................................. 3
2.3 Lymphocyte viability.......................................................................................................... 3
2.4 Selection of oil samples ...................................................................................................... 3
2.5 Measurement of cytokine release........................................................................................ 4
3
Results .......................................................................................................................................... 5
3.1 Effect of emulsified oil on lymphocyte viability ................................................................ 5
3.2 Mitogen stimulation of cytokine release............................................................................. 5
3.3 Effect of emu oil on cytokine release ................................................................................. 6
4.
Discussion..................................................................................................................................... 7
5.
References .................................................................................................................................... 8

Executive Summary
What the report is about & who it is targeted at
To maximise the market potential for emu oil, the Australian Emu Industry needs to overcome two
problems. The first is to identify those factors that result in the different levels of its antiinflammatory activity, thereby allowing the consistent production of oil with known anti-inflammatory
efficacy. The second problem is to develop a sensitive in vitro assay to measure the anti-inflammatory
activity of each batch of oil to grade its potential anti-inflammatory efficacy.
The inflammatory reaction involves the release of a range of inflammatory mediators (cytokines) from
lymphocytes. It is hypothesised, that as an anti-inflammatory agent, emu oil will suppress the
production of these cytokines from human lymphocytes. It is further hypothesised that the amount of
suppression of the cytokine mediators of the inflammatory response from activated human T
lymphocytes will directly correlate with the anti-inflammatory activity of individual oil samples. This
report discusses the work undertaken to investigate these two hypotheses.

Aims & methodology

The aim of this project was to develop an in vitro assay to provide a quantitative measure of the effect
of emu oil on the production of cytokines from human lymphocytes. To develop this assay the
following specific aims were addressed.

Establish a concentration of emulsified emu oil in fetal calf serum that is non-toxic to the
cultured human lymphocytes
Examine the effect of emu oil on the in vitro production of inflammatory mediators from
stimulated human lymphocytes.
Determine whether different batches of emu oil differentially suppress the in vitro production
of inflammatory mediators from stimulated human lymphocytes.

Implications & recommendations

Emu oil at 1% v/v was found to form a stable emulsion when sonicated in 10% fetal calf
serum. Furthermore this emulsion (at 1%) proved to be non-toxic to human lymphocytes.
Emu oil, when applied to stimulated human lymphocytes altered the production of cytokine
inflammatory mediators.
Different batches of emu oil had different effects on the production of individual cytokines.

This study demonstrated it is possible to form a stable emulsion of emu oil that is non-toxic to human
lymphocytes. Emu oil was found to alter the production of pro-inflammatory cytokines from activated
human lymphocytes in vitro. Furthermore individual oils had different effects on the production of the
cytokines. This finding of suppression of pro-inflammatory cytokines from activated human
lymphocytes supports animal studies which demonstrated that emu oil has anti-inflammatory activity
when applied topically. In addition, that individual oils can either suppress or stimulate specific
cytokines to different extents supports the possibility that different emu oils may have different antiinflammatory efficacy. Therefore, the in vitro assay developed in this short-term study shows promise
as a measure of the potential anti-inflammatory efficacy of individual oils. Refinement of the in vitro
assay is however necessary to be able to definitively quantify the effects of emu oil on the production
of the cytokines. These refinements are given under 'Further work'. Finally, to be able to use this in
vitro assay as a measure of the potential anti-inflammatory efficacy of emu oil it is necessary to
correlate the effect of each oil on cytokine production from activated human lymphocytes to its ability
to suppress inflammation in vivo.

vi

Further work
Further refinement of the in vitro assay is neccessary to establish a quantitative measure of the effect
of individual emu oil samples on cytokine release from activated human lymphocytes. These
refinements are tabulated below.

Increase the number of emu oils tested and conduct the in vitro assay in triplicate for each oil
to provide adequate samples for statistical analysis of the data.
Determine the toxicity of the mitogen (DHA) on lymphocyte viability so that the effect of the
each emulsified oil sample on cytokine production can be definitively quantitated.
Reduce the variation in the known concentration of emu oil in the emulsion by using pipette
tips especially designed for viscous fluids.
Assess the potential toxicity of emu oil between 1 and 2% (v) on lymphocyte viability.
Determine whether emu oil has a dose-related effect on production of the different cytokines.
This is necessary to be able to determine whether the effect on cytokine production is a doserelated effect of the emulsified oil or is a result of inherent unique properties of the individual
oils.
To validate this in vitro assay as a measure of the potential anti-inflammatory efficacy of emu
oil, it is necessary to correlate the effect of each oil on cytokine production from activated
human lymphocytes to its ability to suppress inflammation in vivo.

vii

1. Introduction
1.1

Background and overall aims

To date scientific evidence for the anti-inflammatory activity of emu oil has been largely confined to
animal models of inflammation. Topical application of emu oil has been reported to reduce
inflammation in mice (Lpez et al. 1999; Yoganathan et al. 2003) and rats (Snowden & Whitehouse
1997). Furthermore, oils sourced from emus from different habitats and rendered by different
processes vary in their anti-inflammatory efficacy when applied to the skin of rats (Snowden &
Whitehouse 1997; Whitehouse et al. 1998).
The overall aim of this short term study is to refine an in vitro test to provide a quantitative measure of
the potential anti-inflammatory efficacy of different batches of emu oil. Subsequent work will then
investigate factors that may influence the anti-inflammatory activity of emu oil using the in vitro assay
as a measure of its potential anti-inflammatory efficacy. Factors to be investigated in future work will
include the age and sex of the birds at the time of slaughter as well as different rendering processes of
the oil.
The inflammatory reaction involves the release of a range of inflammatory mediators from
lymphocytes. These include the cytokines interleukin (IL)-8, IL-1, IL-6, IL-10, IL-12p70 and
tumour necrosis factor- (TNF-). It is hypothesised, that as an anti-inflammatory agent, emu oil will
suppress the production of these cytokines from activated human T lymphocytes. We further
hypothesise that the amount of suppression of the cytokine mediators of the inflammatory response
from activated human T lymphocytes will directly correlate with the anti-inflammatory activity of
individual oil samples.
To develop an in vitro assay, it is essential that the oil is emulsified in a medium that is not toxic to the
human T lymphocytes. In preliminary studies in our laboratory at Flinders University we have been
able to emulsify the oil samples by sonication in culture medium containing 10% fetal calf serum at
37C. The current work examines the effect of emu oil on the viability of human T lymphocytes and
cytokine production with mitogen activation.

1.2

1.3

Specific aims
Establish a concentration of emulsified emu oil in fetal calf serum that is non-toxic to the
cultured human lymphocytes.
Examine the effect of emu oil on the in vitro production of inflammatory mediators from
stimulated human lymphocytes.
Determine whether different batches of emu oil differentially suppress the in vitro production
of inflammatory mediators from stimulated human lymphocytes.

Emu oil industry perspective

The Australian Emu Industry needs to develop its products to maintain its sustainability. Currently
income is received from the skins, meat and oil. Due to the high costs of both transport to the abattoir
and subsequent processing of the emus it is essential to maximise profits from the sale of each of the
skins, meat and oil.

To maximise the market potential for emu oil, the Industry needs to overcome two problems. The first
is to identify those factors that result in the different levels of its anti-inflammatory activity, thereby
allowing the consistent production of oil with known anti-inflammatory efficacy. The second problem
is to develop a sensitive in vitro assay to measure the anti-inflammatory activity of each batch of oil to
grade its potential anti-inflammatory efficacy.
Market research to date has highlighted the commercial potential of emu oil in pharmaceutical (high
grade) and cosmetic (lower grade) applications. Assuming 5% market penetration by emu oil
indicates a demand for over 750,000 litres of oil per annum. At $70/litre this would result in sales of
Australian product in excess of $50 million per annum. Initial indications are that oil with high antiinflammatory efficacy would command a price of at least $200 per litre on a world market. This
compares to the current market price of $50-70 per litre for emu oil with relatively low antiinflammatory levels.
Enhancing the anti-inflammatory levels of emu oil raises its potential to compete with non-specific
anti-inflammatory drugs (NSAIDs) that can have severe side effects. Indications are that the market
NSAIDs in the treatment of arthritis in major developed countries is in excess of US$10 billion per
annum. It is reasonable to expect that a successful natural alternative with no untoward side effects
could target a market share of 2% approximating US$200 million. The information on market
research was kindly provided by the Industry partners.

2. Methods
2.1

Emulsification of the emu oil

Oil samples (1ml) were sonicated for 20 seconds in RPMI-1460 medium (SAFC BiosciencesTM)
containing 10% fetal bovine serum (Gibco), 13mM NaHCO3, 2mM glutamine, 300 g/ml
streptomycin and 8 g/ml penicillin (complete RPMI medium). The emulsified oil samples were
maintained at 37C. Emulsions were prepared in duplicate at concentrations of 1%, 0.1%, 0.01% and
0.001% v/v. As the oil was extremely viscous care was used to pipette the initial pure oil into the fetal
calf serum.
Initial studies in our laboratory examined the effects of different concentrations of -cyclodextrin, cyclodextrin or -cyclodextrin (0.1, 1% and 10%) on emulsification of the emu oil. As the oil was
found to form a stable emulsion with sonication in the RPMI-1460 medium in the absence of any of
the cyclodextrins, they were excluded from future preparations.

2.2

Lymphocyte source

Two different sources of lymphocytes were used in this study. These were the Jurkat lymphocyte line
and human peripheral blood lymphocytes. The Jurkat cell line was chosen as it is maintained in the
Department of Immunology, Allergy & Arthritis at Flinders Medical Centre and thus provided an
unlimited source of cells. Jurkat is a human T lymphocyte cell line that has been transformed to
survive in culture. Human peripheral blood lymphocytes were separated from whole blood by density
gradient separation using Ficoll PaqueTM Plus (GE Healthcare). Blood was taken from healthy
volunteers by venipuncture (FMC ethics approval FcREC 067/78.25).

2.3

Lymphocyte viability

For in vitro testing of the different concentrations of the oil on lymphocyte viability, 0.5ml of each oil
sample was incubated overnight at 37C in 5% CO2, with 105 cells/ml in 0.5ml of the complete RPMI
medium. All samples were prepared in duplicate. At the end of the incubation period the lymphocytes
were centrifuged at 500g for 5 minutes to pellet the cells. The cells were then resuspended and
incubated for 15 minutes at room temperature in 100ml of 10% fetal calf serum to which was added
7.5M propidium iodide (ICN Biochemicals). Viability of the lymphocytes was analysed by
propidium iodide exclusion on a BD FACSCanto flow cytometer. In the initial experiments, designed
to determine the effect of the oil on lymphocyte viability, the different concentrations of oil were
prepared from aliquots of the same sample.

2.4

Selection of oil samples

In the second set of experiments, the incubation medium contained 1% v/v emulsified oil, with the
exception of one incubation medium that contained 2% v/v of one of the oil samples. The emulsified
samples were prepared from four different batches of emu oil sourced from different emus and
prepared by varying rendering processes. These oils were chosen in an attempt to maximise the
chance of the potential different anti-inflammatory activity of the samples.
Due to restraints of both time to conduct the project and the number of samples that could be analysed
with the cytokine assay kit available, the number of samples that could be examined was limited.
Consequently it was only possible to emulsify one oil sample in duplicate. In addition, one of the oils
(coded #5) was selected at random from the four batches and prepared at 1% (5a) and 2% v/v (5b).
This was to determine whether the higher concentration of the oil had a greater effect on inhibiting the
production of any of the cytokines compared to 1% v/v of emulsified oil. All the emulsified oil

samples were coded prior to analysis and incubations were performed in duplicate. The four different
batches of oil were coded #1, #4, #5, #6 and #12, where #s 1and 12 were taken from the same batch of
oil.

2.5

Measurement of cytokine release

Prior to incubation, lymphocytes were stimulated with the mitogens phytohemagglutinin 0.1ng/ml
PHA; Sigma) or 10ng/ml phorbol 12-myristate 13-acetate (PMA; Sigma). These were added to the
medium to stimulate cytokine release into the supernatant from the lymphocytes. The effect of the
mitogens on cytokine release was examined in separate experiments on lymphocytes obtained from
both the Jurkat cell line and those separated from whole blood. Cultured lymphocytes were incubated
with either both or only one of the mitogens.
After culturing the cells overnight, the supernatant was retrieved by centrifugation and assayed for IL8, IL-1, IL-6, IL-10, TNF- and IL-12p70 using a BD Cytometric Bead Array (CBA) Human
Inflammation Kit (BD Biosciences, USA). The amount of each cytokine in the culture supernatant
was measured by the intensity of fluorescence using by a BD FACSCanto flow cytometer

3
3.1

Results
Effect of emulsified oil on lymphocyte viability

The Jurkat cells (human T lymphocyte cell line) and those separated from whole blood, remained
viable at all concentrations (1%, 0.1%, 0.01% and 0.001%v/v) of the oil after incubation. As 1% oil
v/v proved to be non-toxic to the lymphocytes this concentration was used in all future experiments.

3.2

Mitogen stimulation of cytokine release

The Jurkat cells showed only a minimal increase in the level of IL-8 production in response to either
the phorbol ester (PMA) or PHA compared to the unstimulated cells. No increase in the levels of any
other cytokines was observed with stimulation with either mitogen. This low level of cytokine release
from the Jurkat cells rendered them inappropriate for testing the effects of emu oil on cytokine release.
Unstimulated peripheral blood lymphocytes naturally secreted IL-8 and IL-6 at high levels whereas
IL-1 and TNF were released at intermediate levels. The unstimulated cells showed no release of
either IL-10 or IL-12p70. Stimulation with PHA resulted in a marked increase in the levels of all the
cytokines IL-1, IL-6, IL-10, IL-8 and TNF-, except IL-12p70 which was not released with PHA
stimulation (Figure 3.1). A small decrease in cell viability was observed with PHA as there was an
increase in the percentage of necrotic cells compared to the untreated cells. Stimulation with PMA +/PHA resulted in a reduction in cytokine release compared to stimulation with PHA alone. This effect
may be due to higher toxicity of the PMA compared to PHA resulting in cell death. Consequently
stimulation of the peripheral blood lymphocytes by PHA was used in all future experiments.

Untreated cells
PHA stimulated cells
Figure 3.1 Scatter plots showing the effects of PHA stimulation on cytokine production from human
peripheral blood lymphocytes. The axes show mean fluorescence intensity, where B-585/42 is Rphycoerythrin and B-670LP is fluorescein isothiocyanate. An increase in cytokine production is seen
by a right shift of the fluorescent markers with PHA stimulation.

3.3

Effect of emu oil on cytokine release

Cells stimulated with PHA and additionally cultured with the 1% oil, showed an increase in cell
viability compared to PHA treatment alone. All four batches of the 1% v/v oil had variable effects on
cytokine levels (Table 3.1). All the oils suppressed PHA-stimulation of IL-10 production. With the
exception of suppression of IL-10 levels, two of the oils, coded #1(#12) and #6 had no effect on the
production of IL-8, IL-1 or IL-6, although production of TNF- increased by approximately 30%. A
similar response on cytokine production was observed with the duplicate oil of the same batch as #1
(coded #12). In contrast, oils numbered 4 and 5 (shown in Figure 3.2), sampled from different batches,
decreased the production of IL-1, IL-6, and TNF-. Levels of each of these three cytokines in the
culture supernatant decreased by greater than 50% with oil coded #5.
Increasing the concentration of the oil from 1% to 2% v/v almost totally suppressed the levels of all
cytokines. A greater percentage of the cells appeared to be necrotic compared to the cells similarly
incubated in 1% oil although this effect was definitively quantitated.
As IL-12p70 production remained low with PHA stimulation, the effect of the oil on this cytokine
could not be determined.

Table 3.1

Mean fluorescent intensities of cytokines in the culture supernatant following

treatment with emu oil

Treatment

IL-8

IL-1

IL-6

IL-10

TNF-

IL-12p70

1% Oil (no cells/ no PHA)

Untreated cells
PHA-treated cells
1% Oil +cells (no PHA)
1% Oil #1 + PHA
1% Oil #4 + PHA
1% Oil #5a + PHA
2% Oil #5b + PHA
1% Oil #6 + PHA
1% Oil #12 + PHA

978
109877
146545
141304
149660
139561
105961
60
128624
131725

58
5766
41391
14651
44381
3772
1112
61
39623
38579

287
69277
166727
142794
171357
148591
72962
57
163999
161470

33
688
11883
495
3863
278
334
38
989
2359

92
5258
34485
29861
40652
21855
6238
64
40421
41618

65
248
221
133
259
277
330
65
222
207

PHA stimulated cells

PHA stimulated cells + oil

Figure 3.2 Scatter plots showing effects of 1% oil v/v on cytokine production with PHA-stimulation
of peripheral lymphocytes. A reduction in cytokine levels is seen by a left shift of the fluorescent
markers following incubation with the emu oil.

4. Discussion
Two of the four oils suppressed the production of the pro-inflammatory cytokines IL-1, IL-6, and
TNF-. The suppression of these pro-inflammatory cytokines is consistent with a study using a mouse
model of inflammation, which reported a decrease in both IL-1 and TNF- following topical
application of emu oil (Yoganathan et al. 2003). Yoganathan and colleagues also reported that the
decrease in these pro-inflammatory cytokines was associated with a decrease in inflammation. In
addition, different preparations of emu oil were found to vary in their anti-inflammatory activity
against adjuvant-induced arthritis in rats (Snowden & Whitehouse 1997; Whitehouse et al. 1998). A
plausible hypothesis, taking into account these reports in the literature together with the data from the
present study, is that the variable ability of the different oils to suppress cytokine production from the
stimulated lymphocytes may be related to differences in the anti-inflammatory activity between the
oils. However, whilst the remaining two oils had no apparent affect on the levels of the proinflammatory cytokines IL-8, IL-1, IL-6, the production of TNF- increased by approximately 30%.
This increased production of TNF- complicates any extrapolation of the cytokine levels in the assay
to the potential anti-inflammatory activity of the oil.
In our pilot study only four different oils were tested and a single oil was emulsified in duplicate.
Approximately a 15% variability in cytokine production was observed between the two preparations.
Although care was taken to pipette the viscous oil into the fetal calf serum, it is possible that some
variation in the levels of cytokines detected in the supernatant was due to slightly different amounts of
oil in the initial emulsion. Another factor that may have affected the levels of the cytokine levels in
the culture supernatant is the viability of the mitogen-stimulated lymphocytes. PHA did reduce
lymphocyte viability, although cells stimulated with PHA and additionally cultured with the 1% oil,
showed an increase in cell viability compared to PHA treatment alone. It is therefore possible that at a
concentration of 1% the oil may have exerted a protective effect against PHA-induced injury.
Treatment of the PHA-stimulated lymphocytes with 2% emu oil (v/v) almost totally suppressed the
levels of all cytokines. One explanation for this effect is that a constituent(s) of emu oil is suppressing
cytokine production. In support of this hypothesis, this particular oil, selected at random prior to
incubation, markedly reduced production of IL-1, IL-6, and TNF- at 1% (v/v). The possibility
cannot be discounted however, that the higher concentration of the oil may have been toxic to the
cells. In support of this, although not definitively quantitated, a greater percentage of the cells
appeared to be necrotic compared to the cells similarly incubated in 1% oil. Further work is required
to assess the potential toxicity of emu oil between 1 and 2% (v/v) on lymphocyte viability and to
determine whether emu oil has a dose-related effect on production of the different cytokines.
In summary, emu oil at 1% v/v was found to form a stable emulsion when sonicated in 10% fetal calf
serum. Furthermore this emulsion (at 1%) proved to be non-toxic to human lymphocytes. Emu oil,
when applied to PHA-stimulated human lymphocytes altered the production of cytokine inflammatory
mediators. Different batches of emu oil had variable effects of cytokine production. Further work is
required to develop this assay which promises to provide a rapid and quantitative measure of the effect
of emu oil on cytokine production.

5. References
Lpez, A, Sims ED, Ablett, RF, Skinner, RE, Lger, LW, Lariviere CM, Jamieson, LA, MartnezBurnes, J & Zawadzka, GG, 1999, Effect of emu oil on auricular inflammation induced with croton
oil in mice, American Journal of Veterinary Research, vol. 60(12), pp. 1158-1611.
Snowden, JM & Whitehouse, MW, 1997, Anti-inflammatory activity of emu oil in rats,
Inflammopharmacology, vol. 5, pp 127-132.
Whitehouse, MW, Turner AG, Davis CKC Roberts, MS, Emu oils(s): A source of non-toxic
transdermal anti-inflammatory agents in aboriginal medicine, Inflammopharmacology, vol. 6, pp 1-8.
Yoganathan, S, Nicolosi, R, Wilson, T, Handelman, G, Scollin, P, Tao, R, Binford, P & Orthoefer, F,
2003, Anatogonism of croton oil inflammation by topical emu oil in CD-1 mice, Lipids, vol. 38, pp.
603-607.